Human Tumor Necrosis Factor alpha ELISA Kit

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Descriere

Specificații

Brand

Antibodies

Mărimi disponibile

96T · 192T · 192T x 3

Țintă

Tumor Necrosis Factor alpha

Applications

Sandwich ELISA

Reactivity

Human

Assay type

Sandwich (quantitative)

Sensitivity

< 4 pg/ml

Sample type

Serum, plasma, cell culture supernatant, and other biological fluids.

Cross reactivity

This assay has shown no cross-reactivity with other cytokines tested.

Detection type

Colorimetric

Platform

Pre-coated Microplate (12 x 8 well strips)

Product range

4-2000 pg/ml

Assay Time

2h 30m - 3h

Specificitate

This sandwich ELISA recognises both natural and recombinant human TNF-a.

Principiul testului

This TNF-a enzyme linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to TNF-a. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated monoclonal antibody preparation specific for TNF-a and incubated. TNF-a, if present, will bind and become immobilized by the antibody pre-coated on the wells and then be “sandwiched” by biotin conjugate. The microtiter plate wells are thoroughly washed to remove unbound TNF-a and other components of the sample. In order to quantify the amount of TNF-a present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Avidin is a tetramer containing four identical subunits that each has a high affinity-binding site for biotin. The wells are thoroughly washed to remove all unbound HRP-conjugated Avidin and a TMB (3,3'5,5' tetramethyl-enzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain TNF-a, biotin conjugated antibody, and enzyme-conjugated Avidin will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the colour change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. In order to measure the concentration of TNF-a in the samples, this kit includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus TNF-a concentration (pg/mL). The concentration of TNF-a in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Storage

Shipped and stored at 4°C.